780 CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(4) This serum should be very clear, 2. Meat extract . la . 



lZ77r: -^^*-"-^^pa^e 3. Peptone (Witte). ; •.:::: ;:::; Zt 



(5) Dissolve 10 to 1.5 g. sodium salt of 5.' Caliumacid malate 60 f ' 



nucleic acid (Bohrenger) in 200.0 cc. 6. Glucose .J' 



of sterile (4) by shaking. 7. ge.um (beef) '"'^^ 



(6) Pass carbonic acid from a Kipp ap- 8. Potassium tellurite (1 0% 

 paratus thru (5) for 2 or 3 minutes. solution) 



This tends to clear the medium. pr„„- .■ 



(7) Distribute into tall test tubes about (T) D 's^iU 004 .,- , 

 f full and heat in a water bath on 2 rZ ' '° 

 three successive days for one hour m Ar\r} I'no/ 1 



each day at 60°C. ^f, f ^^ ^-^^^ g^^«««^- 



(8) After removal from the water bath ^^ t . ^^'^ ^^^ *° ^ ^^"^' ^'''^ '*""^^ 

 place into ice cold water. ... t^"" inn'o""' , , • 



(9) At the time of the fourth sterilization ^^ l" ?•" '''t '^^^*"'' ^^^ ^'^ 

 solidify the serum by gradually g'j^.f "' "^^^ Potassium tellurite 

 raising the temperature to 70 °C fr,\ d^ ^ ^°^" 



You can obtain three different degrees ^^ '"^ ^^*" "^'^^^^ ^^^^ bibulous 



of hardness of the serum. By heating Paper mside covers. 



at a low temperature one obtains a ^'^^ Solidify at 85-90°C. 



soft, clear and transparent medium. Sterilization: Sterilize by repeated heating 



By heating at a high temperature one (temperature not given), 



obtains a hard solid nearly opaque ^^e: Isolation of diphtheria bacilli, 



medium. Then there is a medium Author reported that diphtheria colonies 



between the two extremes. were coal black due to the reduction of 



Sterilization: Sterilize (5) by the fractional tellurium. 



method in the steamer. Further sterili- Reference: Conradi and Troch (1912 pp 



zation is obtained in step (7) under the 1652-1653). 



preparation. 



Use: Cultivation of spirochetes. 2414. Hall and Stone's Glycerol Serum 



Variants : Futaki, et al., studied the motility Medium 



of Spirochaeta morsus niuris (cause of rat 



bite fever) on a medium prepared as Constituents: 



follows: 1- Water 1000.0 cc. 



(1) Shake 0.5 to 0.75 g. of sodium nucleate ^- ^^Ptone (Witte) 10.0 g. ' 



with 100.0 cc. horse serum until the ^- ^^^^ 5.0 g. 



nucleate has dissolved. *■ Glucose lO.o g. 



(2) Pass CO2 thru the medium for 2 or 3 ^- ^^y^^rol 5O.0 g. 



minutes until the medium is trans- ^' ®^^"™' blood 3000.0 cc. 



parent. Preparation: 



(3) Heat on three successive days for ^^^ Dissolve 2, 3, 4 and 5 in 1. 



about an hour at 60°C. (2) Mix one part of (1) with 3 parts of 6. 



(4) On the 4th day heat to 65°C. for 30 ^^^ Filter thru Berkefeld filter, 

 minutes, when a fluid and coagulated ^^^ ^ube. 



portion is formed. (5) Slant. 

 References: Shmamine (1912 p. 313), Futaki, Sterilization: Sterilize in autoclave at 10 



Takaki, Taniguchi and Osumi (1917 p. pounds pressure for 30 minutes. 

 ^'- Use: Cultivation of diphtheroid bacillus 



2413. Conradi and Troch's Extract Broth °f P^eisz-Nocard from equine and bovine 



Serum Medium abscesses. Author reported that several 



^ ^. strains of B. diphtheriae will grow on 



Constituents: this medium. 



^' ^^^^^'^ • • 1000-0 cc. Reference: Hall and Stone (1916 p. 196). 



